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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Membrane-associated GAlP is a phosphoprotein and can be phosphorylated by clathrin-coated vesicles
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Membrane-associated GAlP is a phosphoprotein and can be phosphorylated by clathrin-coated vesicles

机译：膜相关的GAlP是一种磷蛋白，可以被网格蛋白包被的囊泡磷酸化

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摘要

GAIP (G αinteracting protein) is a member of the RGS (regulators of G protein signaling) family and accelerates the turnover of GTP bound to Goi, Gαq. and Gα13. There are two pools of GAlP--a soluble and a membrane-anchored pool. The membrane-anchored pool is found on clathrin-coated vesicles (CCVs) and pits in rat liver and AtT-20 pituitary cells. By treatment of a GAlP-enriched rat liver fraction with alkaline phosphatase, we found that membrane- bound GAlP is phosphorylated. By immunoprecipitation carried out on [~32Plorthophosphate-labeled AtT-20 pituitary cells stably ex- pressing GAlP, ~32P-labeling was associated exclusively with the membrane pool of GAlP. Phosphoamino acid analysis revealed that phosphorylation of GAlP occurred largely on serine residues. Re- combinant GAlP could be phosphorylated at its N terminus with purified casein kinase 2 (CK2). It could also be phosphorylated by isolated CCVs in ds. Phosphorylation was Mn~2+-dependent, using both purified CK2 and CCVs. Ser-24 was identified as one of the phosphorylation sites. Our results establish that GAlP is phos- phorylated and that only the membrane pool is phosphorylated, suggesting that GAlP can be regulated by phosphorylation events taking place at the level of clathrin-coated pits and vesicles.
机译：GAIP（Gα相互作用蛋白）是RGS（G蛋白信号调节剂）家族的成员，可加速与GoiGαq结合的GTP的转换。和Gα13。有两个GAlP池-可溶池和膜锚定池。膜锚定池在网格蛋白包被的囊泡（CCV）和大鼠肝脏和AtT-20垂体细胞的凹坑中发现。通过用碱性磷酸酶处理富含GAlP的大鼠肝脏部分，我们发现膜结合的GAlP被磷酸化了。通过在稳定表达GAlP的[〜32焦磷酸盐标记的AtT-20垂体细胞上进行的免疫沉淀，〜32P标记仅与GAlP的膜池相关。磷酸氨基酸分析表明，GAlP的磷酸化主要发生在丝氨酸残基上。重组的GA1P可以在其N末端被纯化的酪蛋白激酶2（CK2）磷酸化。它也可能在ds中被孤立的CCV磷酸化。使用纯化的CK2和CCV，磷酸化均依赖于Mn〜2 +。 Ser-24被鉴定为磷酸化位点之一。我们的研究结果表明，GAlP被磷酸化了，而只有膜池被磷酸化了，这表明GAlP可以通过在网格蛋白包被的凹坑和囊泡水平发生的磷酸化事件来调节。

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